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杂交捕获的力量 - 用100个死后样本的扩展基因面板来说明,重点是突然无法解释的死亡

Daniel Kling¹, Emma Adolfsson², Henrik Gréen³

¹Department of Forensic Genetics and Forensic Toxicology, National Board of Forensic Medicine, Linköping, Sweden.

Forensic science international. Genetics

|October 22, 2024

概括

法医分子尸检使用2422个基因的面板来确定突然意外死亡 (SUD) 的遗传原因. 这种先进的杂交捕获方法改善了测序,并有效地优先考虑研究的遗传变异.

科学领域:

法医遗传学法医遗传学
分子尸体解剖学分子尸体解剖学
基因组医学是基因组医学.

背景情况:

突然意外死亡 (SUD) 通常具有潜在的遗传基础.
使用DNA测序的分子尸检对于在医学研究结果不确时确定死亡原因至关重要.
目前的方法在基因面板大小和测序范围上有所不同,从有限的基因面板到整个外基因组测序.

研究的目的:

为了评估2422基因向捕获面板的性能,用于法医分子解剖.
在NextSeq 550和MiSeq FGx平台上评估测序成功指标.
开发和验证一个逐步程序来优先考虑与SUD相关的遗传变异.

主要方法:

使用Twist Bioscience的双链DNA探针的杂交捕获方法被用来准与SUD相关的2422个基因.
从有史以来的法医病例中获得的98个尸体样本,其中死亡原因未确定,以及之前的负分子解剖,进行了测序.
在NextSeq 550和MiSeq FGx平台上评估性能,重点关注覆盖范围和统一性.

主要成果:

2422个基因组显示出高的捕获统一性 (平均折叠80=1.5),确保了足够的覆盖率 (20X),同时最大限度地降低了测序成本.
MiSeq FGx平台使用2×300bp的测序,改善了覆盖范围,特别是在具有挑战性的地区.
一个逐步的生物信息方法有效地优先考虑了基因变异,确定了很少的 (0-3) 高优先级变异,需要在所有基因层进行手动检查.

结论:

2422基因向捕获面板是法医分子尸检的高性能和低成本工具.
先进的测序策略,如MiSeq FGx上的2×300 bp,提高了复杂遗传分析的数据质量.
开发的生物信息管道有效地过变体,大大减少了识别SUD遗传原因的工作量.

关键词:

法医遗传学法医遗传学分子解剖的方法是分子解剖.突然的无法解释的死亡.有针对性的测序.

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